Hemoglobin and Chapter

Iron Metabolism 11
„ INTRODUCTION
„ NORMAL HEMOGLOBIN CONTENT
„ FUNCTIONS
„ STRUCTURE
„ TYPES OF NORMAL HEMOGLOBIN
„ ABNORMAL HEMOGLOBIN
„ ABNORMAL HEMOGLOBIN DERIVATIVES
„ SYNTHESIS
„ DESTRUCTION
„ IRON METABOLISM

„ INTRODUCTION „ FUNCTIONS OF HEMOGLOBIN

Hemoglobin (Hb) is the iron containing coloring matter of „ TRANSPORT OF RESPIRATORY GASES
red blood cell (RBC). It is a chromoprotein forming 95%
of dry weight of RBC and 30% to 34% of wet weight. Main function of hemoglobin is the transport of respiratory
Function of hemoglobin is to carry the respiratory gases, gases:
oxygen and carbon dioxide. It also acts as a buffer. 1. Oxygen from the lungs to tissues.
Molecular weight of hemoglobin is 68,000. 2. Carbon dioxide from tissues to lungs.

„ NORMAL HEMOGLOBIN CONTENT 1. Transport of Oxygen

When oxygen binds with hemoglobin, a physical process
Average hemoglobin (Hb) content in blood is 14 to 16 g/dL.
called oxygenation occurs, resulting in the formation of
However, the value varies depending upon the age and
oxyhemoglobin. The iron remains in ferrous state in this
sex of the individual.
compound. Oxyhemoglobin is an unstable compound
and the combination is reversible, i.e. when more oxygen
Age
is available, it combines with hemoglobin and whenever
At birth : 25 g/dL oxygen is required, hemoglobin can release oxygen
After 3rd month : 20 g/dL readily (Chapter 125).
After 1 year : 17 g/dL When oxygen is released from oxyhemoglobin, it is
From puberty onwards : 14 to 16 g/dL called reduced hemoglobin or ferrohemoglobin.
At the time of birth, hemoglobin content is very high
because of increased number of RBCs (Chapter 9). 2. Transport of Carbon Dioxide
When carbon dioxide binds with hemoglobin, carbhe­
Sex
moglobin is formed. It is also an unstable compound
In adult males : 15 g/dL and the combination is reversible, i.e. the carbon dioxide
In adult females : 14.5 g/dL can be released from this compound. The affinity of
 78 Section 2 t Blood and Body Fluids

hemoglobin for carbon dioxide is 20 times more than to 12th week after birth. Both the types of hemoglobin
that for oxygen (Chapter 125). differ from each other structurally and functionally.

„ BUFFER ACTION Structural Difference

Hemoglobin acts as a buffer and plays an important role In adult hemoglobin, the globin contains two α-chains
in acid­base balance (Chapter 5). and two β-chains. In fetal hemoglobin, there are two α
chains and two γ-chains instead of β-chains.
„ STRUCTURE OF HEMOGLOBIN
Functional Difference
Hemoglobin is a conjugated protein. It consists of a
protein combined with an iron­containing pigment. The Functionally, fetal hemoglobin has more affinity for
protein part is globin and the iron­containing pigment oxygen than that of adult hemoglobin. And, the oxygen­
is heme. Heme also forms a part of the structure of hemoglobin dissociation curve of fetal blood is shifted to
myoglobin (oxygen­binding pigment in muscles) and left (Chapter 125).
neuroglobin (oxygen­binding pigment in brain).
„ ABNORMAL HEMOGLOBIN
„ IRON
Abnormal types of hemoglobin or hemoglobin variants
Normally, it is present in ferrous (Fe2+) form. It is in
are the pathologic mutant forms of hemoglobin. These
unstable or loose form. In some abnormal conditions,
variants are produced because of structural changes
the iron is converted into ferric (Fe3+) state, which is a
in the polypeptide chains caused by mutation in the
stable form.
genes of the globin chains. Most of the mutations do not
„ PORPHYRIN produce any serious problem. Occasionally, few muta­
tions result in some disorders.
The pigment part of heme is called porphyrin. It is There are two categories of abnormal hemoglobin:
formed by four pyrrole rings (tetrapyrrole) called, I, II, III 1. Hemoglobinopathies
and IV. The pyrrole rings are attached to one another by 2. Hemoglobin in thalassemia and related disorders.
methane (CH4) bridges.
The iron is attached to ‘N’ of each pyrrole ring and ‘N’ 1. Hemoglobinopathies
of globin molecule.
Hemoglobinopathy is a genetic disorder caused by
„ GLOBIN abnormal polypeptide chains of hemoglobin.
Some of the hemoglobinopathies are:
Globin contains four polypeptide chains. Among the four i. Hemoglobin S: It is found in sickle cell anemia.
polypeptide chains, two are chains and two are α-chains In this, the α-chains are normal and β-chains are
(Refer Table 11.1 for molecular weight and number of abnormal.
amino acids in the polypeptide chains).
ii. Hemoglobin C: The β-chains are abnormal. It is
TABLE 11.1: Molecular weight and number of amino found in people with hemoglobin C disease, which
acids of polypeptide chains of globin is characterized by mild hemolytic anemia and
Molecular splenomegaly.
Polypeptide chain Amino acids
weight iii. Hemoglobin E: Here also the β-chains are abnormal.
α-chain 15,126 141 It is present in people with hemoglobin E disease
which is also characterized by mild hemolytic
β-chain 15,866 146
anemia and splenomegaly.
iv. Hemoglobin M: It is the abnormal hemoglobin
„ TYPES OF NORMAL HEMOGLOBIN
present in the form of methemoglobin. It occurs
Hemoglobin is of two types: due to mutation of genes of both in α and β chains,
1. Adult hemoglobin – HbA resulting in abnormal replacement of amino acids.
2. Fetal hemoglobin – HbF It is present in babies affected by hemoglobin M
Replacement of fetal hemoglobin by adult hemoglobin disease or blue baby syndrome. It is an inherited
starts immediately after birth. It is completed at about 10th disease, characterized by methemoglobinemia.
 Chapter 11 t Hemoglobin and Iron Metabolism 79

2. Hemoglobin in Thalassemia and Sources of Carbon Monoxide

Related Disorders
1. Charcoal burning
In thalassemia, different types of abnormal hemoglobins 2. Coal mines
are present. The polypeptide chains are decreased, 3. Deep wells
absent or abnormal. In α-thalassemia, the α-chains are 4. Underground drainage system
decreased, absent or abnormal and in β-thalassemia, 5. Exhaust of gasoline engines
the β-chains are decreased, absent or abnormal 6. Gases from guns and other weapons
(Chapter 14). Some of the abnormal hemoglobins found 7. Heating system with poor or improper ventilation
in thalassemia are hemoglobin G, H, I, Bart’s, Kenya, 8. Smoke from fire
Lepore and constant spring. 9. Tobacco smoking.

Signs and Symptoms of

„ ABNORMAL HEMOGLOBIN DERIVATIVES
Carbon Monoxide Poisoning
‘Hemoglobin derivatives’ refer to a blood test to detect
1. While breathing air with less than 1% of CO, the Hb
and measure the percentage of abnormal hemoglobin
saturation is 15% to 20% and mild symptoms like
derivatives.
headache and nausea appear
Hemoglobin is the only carrier for transport of oxygen, 2. While breathing air with more than 1% CO, the
without which tissue death occurs within few minutes. Hb saturation is 30% to 40%. It causes severe
When hemoglobin is altered, its oxygen carrying symptoms like:
capacity is decreased resulting in lack of oxygen. So, it i. Convulsions
is important to know about the causes and the effects of ii. Cardiorespiratory arrest
abnormal hemoglobin derivatives. iii. Unconsciousness and coma.
Abnormal hemoglobin derivatives are formed by 3. When Hb saturation increases above 50%, death
carbon monoxide (CO) poisoning or due to some drugs occurs.
like nitrites, nitrates and sulphanamides.
Abnormal hemoglobin derivatives are: „ METHEMOGLOBIN
1. Carboxyhemoglobin
2. Methemoglobin Methemoglobin is the abnormal hemoglobin derivative
3. Sulfhemoglobin. formed when iron molecule of hemoglobin is oxidized
Normal percentage of hemoglobin derivatives in total from normal ferrous state to ferric state. Methemoglobin
hemoglobin: is also called ferrihemoglobin.
Carboxyhemoglobin : 3% to 5 % Normal methemoglobin level is 0.6% to 2.5% of total
hemoglobin.
Methemoglobin : less than 3%
Under normal circumstances also, body faces the
Sulfhemoglobin : trace (undetectable). threat of continuous production of methemoglobin. But it
Abnormally high levels of hemoglobin derivates is counteracted by erythrocyte protective system called
in blood produce serious effects. These derivatives nicotinamide adenine dinucleotide (NADH) system,
prevent the transport of oxygen resulting in oxygen lack which operates through two enzymes:
in tissues, which may be fatal. 1. Diaphorase I (nicotinamide adenine dinucleotide
phosphate [NADPH]­dependent reductase): Res­
„ CARBOXYHEMOGLOBIN ponsible for 95% of the action.
2. Diaphorase II (NADPH­dependent methemoglobin
Carboxyhemoglobin or carbon monoxyhemoglobin
reductase): Responsible for 5% of the action.
is the abnormal hemoglobin derivative formed by the
These two enzymes prevent the oxidation of ferrous
combination of carbon monoxide with hemoglobin.
iron into ferric iron.
Carbon monoxide is a colorless and odorless gas. Since
hemoglobin has 200 times more affinity for carbon Methemoglobinemia
monoxide than oxygen, it hinders the transport of oxygen
resulting in tissue hypoxia (Chapter 127). Methemoglobinemia is the disorder characterized by
Normally, 1% to 3% of hemoglobin is in the form of high level of methemoglobin in blood. It leads to tissue
carboxyhemoglobin. hypoxia, which causes cyanosis and other symptoms.
 80 Section 2 t Blood and Body Fluids

Causes of methemoglobinemia level rises above 10 gm/dL, cyanosis occur. Usually,

serious toxic effects are not noticed.
Methemoglobinemia is caused by variety of factors:
1. Common factors of daily life: „ SYNTHESIS OF HEMOGLOBIN
i. Well water contaminated with nitrates and Synthesis of hemoglobin actually starts in proerythro­
nitrites blastic stage (Fig. 11.1). However, hemoglobin appears
ii. Fires in the intermediate normoblastic stage only. Production of
iii. Laundry ink hemoglobin is continued until the stage of reticulocyte.
iv. Match sticks and explosives Heme portion of hemoglobin is synthesized in
v. Meat preservatives (which contain nitrates and mitochondria. And the protein part, globin is synthesized
nitrites) in ribosomes.
vi. Mothballs (naphthalene balls)
vii. Room deodorizer propellants. „ SYNTHESIS OF HEME
2. Exposure to industrial chemicals such as: Heme is synthesized from succinyl­CoA and the glycine.
The sequence of events in synthesis of hemoglobin:
i. Aromatic amines
1. First step in heme synthesis takes place in the mito­
ii. Fluorides
chondrion. Two molecules of succinyl­CoA combine
iii. Irritant gases like nitrous oxide and nitro­
with two molecules of glycine and condense to form
benzene
δ-aminolevulinic acid (ALA) by ALA synthase.
iv. Propylene glycol dinitrate.
2. ALA is transported to the cytoplasm. Two molecules
3. Drugs: of ALA combine to form porphobilinogen in the
presence of ALA dehydratase.
i. Antibacterial drugs like sulfonamides
3. Porphobilinogen is converted into uroporphobilinogen
ii. Antimalarial drugs like chloroquine
I by uroporphobilinogen I synthase.
iii. Antiseptics
4. Uroporphobilinogen I is converted into uroporpho­
iv. Inhalant in cyanide antidote kit
bilinogen III by porphobilinogen III cosynthase.
v. Local anesthetics like benzocaine.
5. From uroporphobilinogen III, a ring structure
4. Hereditary trait: called coproporphyrinogen III is formed by
uroporphobilinogen decarboxylase.
Due to deficiency of NADH-dependant reductase or 6. Coproporphyrinogen III is transported back to the
presence of abnormal hemoglobin M. Hemoglobin M is mitochondrion, where it is oxidized to form proto­
common in babies affected by blue baby syndrome (a porphyrinogen IX by coproporphyrinogen oxidase
pathological condition in infants, characterized by bluish 7. Protoporphyrinogen IX is converted into proto­
skin discoloration (cyanosis), caused by congenital heart porphyrin IX by protoporphyrinogen oxidase.
defect). 8. Protoporphyrin IX combines with iron to form heme
in the presence of ferrochelatase.
„ SULFHEMOGLOBIN
Sulfhemoglobin is the abnormal hemoglobin derivative, „ FORMATION OF GLOBIN
formed by the combination of hemoglobin with hydrogen Polypeptide chains of globin are produced in the
sulfide. It is caused by drugs such as phenacetin or ribosomes. There are four types of polypeptide chains
sulfonamides. namely, alpha, beta, gamma and delta chains. Each
Normal sulfhemoglobin level is less than 1% of total of these chains differs from others by the amino acid
hemoglobin. sequence. Each globin molecule is formed by the
Sulfhemoglobin cannot be converted back into combination of 2 pairs of chains and each chain is made
hemoglobin. Only way to get rid of this from the body is of 141 to 146 amino acids. Adult hemoglobin contains
to wait until the affected RBCs with sulfhemoglobin are two alpha chains and two beta chains. Fetal hemoglobin
destroyed after their lifespan. contains two alpha chains and two gamma chains.

Blood Level of Sulfhemoglobin „ CONFIGURATION

Normally, very negligible amount of sulfhemoglobin is Each polypeptide chain combines with one heme
present in blood which is nondetectable. But when its molecule. Thus, after the complete configuration, each
 Chapter 11 t Hemoglobin and Iron Metabolism 81

FIGURE 11.1: Synthesis of hemoglobin

hemoglobin molecule contains 4 polypeptide chains and Globin is utilized for the resynthesis of hemoglobin.
4 heme molecules. Heme is degraded into iron and porphyrin. Iron is stored
in the body as ferritin and hemosiderin, which are
„ SUBSTANCES NECESSARY FOR reutilized for the synthesis of new hemoglobin. Porphyrin
HEMOGLOBIN SYNTHESIS is converted into a green pigment called biliverdin. In
human being, most of the biliverdin is converted into a
Various materials are essential for the formation of yellow pigment called bilirubin. Bilirubin and biliverdin
hemoglobin in the RBC (Refer Chapter 10 for details). are together called the bile pigments (Details of bile
pigments are given in Chapter 40).
„ DESTRUCTION OF HEMOGLOBIN
After the lifespan of 120 days, the RBC is destroyed „ IRON METABOLISM
in the reticuloendothelial system, particularly in spleen
„ IMPORTANCE OF IRON
and the hemoglobin is released into plasma. Soon, the
hemoglobin is degraded in the reticuloendothelial cells Iron is an essential mineral and an important component
and split into globin and heme. of proteins, involved in oxygen transport. So, human
 82 Section 2 t Blood and Body Fluids

body needs iron for oxygen transport. Iron is important into blood by a protein called ferroportin. In the blood,
for the formation of hemoglobin and myoglobin. Iron is ferric iron is converted into ferrous iron and transported.
also necessary for the formation of other substances
like cytochrome, cytochrome oxidase, peroxidase and „ TRANSPORT OF IRON
catalase. Immediately after absorption into blood, iron combines
with a β-globulin called apotransferrin (secreted by liver
„ NORMAL VALUE AND DISTRIBUTION through bile) resulting in the formation of transferrin.
OF IRON IN THE BODY And iron is transported in blood in the form of transferrin.
Total quantity of iron in the body is about 4 g. Approximate Iron combines loosely with globin and can be released
distribution of iron in the body is as follows: easily at any region of the body.
In the hemoglobin : 65% to 68%
„ STORAGE OF IRON
In the muscle as myoglobin : 4%
As intracellular oxidative heme compound : 1% Iron is stored in large quantities in reticuloendothelial
In the plasma as transferrin : 0.1% cells and liver hepatocytes. In other cells also it is stored
Stored in the reticuloendothelial system : 25% to 30% in small quantities. In the cytoplasm of the cell, iron is
stored as ferritin in large amount. Small quantity of iron
„ DIETARY IRON is also stored as hemosiderin.

Dietary iron is available in two forms called heme and „ DAILY LOSS OF IRON
nonheme.
In males, about 1 mg of iron is excreted everyday through
Heme Iron feces. In females, the amount of iron loss is very much
high. This is because of the menstruation.
Heme iron is present in fish, meat and chicken. Iron in One gram of hemoglobin contains 3.34 mg of iron.
these sources is found in the form of heme. Heme iron Normally, 100 mL of blood contains 15 gm of hemoglobin
is absorbed easily from intestine. and about 50 mg of iron (3.34 × 15). So, if 100 mL of
blood is lost from the body, there is a loss of about 50 mg
Non-heme Iron of iron. In females, during every menstrual cycle, about
50 mL of blood is lost by which 25 mg of iron is lost. This
Iron in the form of nonheme is available in vegetables, is why the iron content is always less in females than in
grains and cereals. Non­heme iron is not absorbed males.
easily as heme iron. Cereals, flours and products of Iron is lost during hemorrhage and blood donation
grains which are enriched or fortified (strengthened) with also. If 450 mL of blood is donated, about 225 mg of iron
iron become good dietary sources of non­heme iron, is lost.
particularly for children and women.
„ REGULATION OF TOTAL IRON IN THE BODY
„ ABSORPTION OF IRON
Absorption and excretion of iron are maintained almost
Iron is absorbed mainly from the small intestine. It is equally under normal physiological conditions. When
absorbed through the intestinal cells (enterocytes) the iron storage is saturated in the body, it automatically
by pinocytosis and transported into the blood. Bile is reduces the further absorption of iron from the gastro­
essential for the absorption of iron. intestinal tract by feedback mechanism.
Iron is present mostly in ferric (Fe3+) form. It is Factors which reduce the absorption of iron:
converted into ferrous form (Fe2+) which is absorbed into 1. Stoppage of apotransferrin formation in the liver, so
the blood. Hydrochloric acid from gastric juice makes that the iron cannot be absorbed from the intestine.
the ferrous iron soluble so that it could be converted 2. Reduction in the release of iron from the transferrin,
into ferric iron by the enzyme ferric reductase from so that transferrin is completely saturated with iron
enterocytes. From enterocytes, ferric iron is transported and further absorption is prevented.